Abstract
Optical nanosensors are based on particles with diameters from 20 to 200 nm containing sensory elements. The latter are comprised of one or more signaling molecules and one or more references, which allow measurements to be ratiometric and hence independent on the amount of sensor. The signaling molecules may range from simple ion-binding fluorophores, e.g., pH-sensitive dyes, to complex biochemical assays. Aptamers are ideal for use in nanosensors because they are relatively easy to modify chemically and hence to transform into signaling molecules, and their binding affinities may be fine-tuned to a desired measuring range in the selection process. Here we first describe the selection of metabolite binding aptamers, how they are transformed into signaling molecules using a molecular beacon construct and then how they are inserted into nanoparticles. Finally, we briefly describe how the sensors are calibrated before inserted into cells to measure metabolite concentration in real time. As examples we present aptamers binding to key metabolites in cells: ATP and fructose 1, 6-bisphosphate (FBP).
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Acknowledgements
This research was supported by EU FP7 Marie Curie program to the Initial Training Network ISOLATE. Tine Daa Schrøder was supported by the Lundbeck Foundation grant to the Nanomedicine Research Center for Cancer Stem Cell-Targeting Therapeutics (NanoCAN).
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Long, Y., Pfeiffer, F., Mayer, G., Schrøder, T.D., Özalp, V.C., Olsen, L.F. (2016). Selection of Aptamers for Metabolite Sensing and Construction of Optical Nanosensors. In: Mayer, G. (eds) Nucleic Acid Aptamers. Methods in Molecular Biology, vol 1380. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3197-2_1
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DOI: https://doi.org/10.1007/978-1-4939-3197-2_1
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3196-5
Online ISBN: 978-1-4939-3197-2
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